Apparent subunit heterogeneity of NAD-dependent glyceraldehyde-3-phosphate dehydrogenase from spinach leaves

Abstract

NAD dependent glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12) from spinach leaves has been previously investigated: the enzyme, homogeneous on conventional chromatography and analytical disc electrophoresis, shows on denaturing condition, non identical subunits of 37,000 and 14,000 daltons which have been separated in equimolar amounts after sodium dodecyl sulphate treatment (Speranza and Gozzer, 1978). Affinity chromatography carried out on these enyzme preparations allowed to separate a fully active enzyme of 150,000 daltons M.W. from a protein of 70,000 daltons M.W. which is devoid of glyceraldehyde-3-phosphate dehydrogenase activity: the purified enzyme shows 37,000 daltons subunits, while the second protein is made up of 14,000 daltons subunits. It is also reported that the 70,000 daltons protein could be separated from the enzyme by selective crystallization in the presence of organic solvents. It is concluded that the NAD-dependent glyceraldehyde-3-phosphate dehydrogenase from spinach leaves, as the homologous enzyme from other sources, is a tetramer of 37,000 daltons subunits.